Monoclonal antibodies and FV specific for CD2 antigen

Abstract

An anti-CD2 monoclonal antibody according to the present invention can be: (1) a chimeric monoclonal antibody CD2 SFv-Ig produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277; (2) a monoclonal antibody having complementarity-determining regions identical with those of CD2 SFv-Ig; or (3) a monoclonal antibody competing with CD2 SFv-Ig for binding to CD2 antigen at least about 80% as effectively on a molar basis as CD2 SFv-Ig. Anti-CD2 monoclonal antibodies according to the present invention, as well as other antibodies that can modulate the interactions between T lymphocytes and monocytes, can be used to inhibit the production of HIV-1 by HIV-1-infected T cells in HIV-1-infected patients. In another use, T cells treated in vitro can be reinfused into AIDS patients to increase the proportion of functional, non-HIV-1-producing T cells in the patient.

We claim: 
     
       1. A single-chain recombinant antibody having specific binding affinity for CD2 antigen, including therein a human constant region sequence including: a hinge region having an amino acid sequence therein of E-P-K-S-S-D-K-T-H-T-S-P-P-S-P (SEQ ID NO:4), the hinge region being located between the variable region sequences and the constant region sequences. 
     
     
       2. The single-chain recombinant antibody of claim 1 wherein the recombinant antibody competes with the single-chain recombinant antibody derived from monoclonal antibody 35.1 and produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277 for binding to CD2 antigen at least as about 90% as effectively on a molar basis as the single-chain recombinant antibody derived from monoclonal antibody 35.1 and produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277. 
     
     
       3. The single-chain recombinant antibody of claim 1 wherein the complementarity-determining regions of heavy chain origin are identical in sequence with those of the single-chain recombinant antibody derived from monoclonal antibody 35.1 and produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277. 
     
     
       4. The single-chain recombinant antibody of claim 1 wherein the complementarity-determining regions of light chain origin are identical in sequence with those of the single-chain recombinant antibody derived from monoclonal antibody 35.1 and produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277. 
     
     
       5. The single-chain recombinant antibody of claim 1 labeled with a detectable marker. 
     
     
       6. The monoclonal antibody of claim 5 wherein the detectable marker is selected from the group consisting of enzymes, paramagnetic materials, members of the avidin-biotin specific binding pair, fluorophores, chromophores, chemiluminophores, heavy metals, and radioisotopes. 
     
     
       7. A composition comprising: (a) a single-chain recombinant antibody of claim 1 in a quantity sufficient to inhibit production of HIV-1 in infected T cells; and   (b) a carrier.   
     
     
       8. The composition of claim 7 wherein the single-chain recombinant antibody is the single-chain recombinant antibody derived from monoclonal antibody 35.1 and produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277. 
     
     
       9. A single-chain recombinant antibody having specific binding affinity for CD2 antigen, including therein a human constant region sequence including: (1) a hinge region having an amino acid sequence therein of E-P-K-S-S-D-K-T-H-T-S-P-P-S-P (SEQ ID NO:4); and   (2) the carboxy-terminal portion of a human constant region with an additional portion of the constant region, and including a linker between light and heavy chain sequences, the hinge region having the amino acid sequence SEQ ID NO:4 being located between the variable region sequences and the constant region sequences, the single chain recombinant antibody being the single-chain recombinant antibody derived from monoclonal antibody 35.1 and produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277.   
     
     
       10. A single-chain recombinant antibody having specific binding affinity for CD2 antigen, including therein a human constant region sequence including: (1) a hinge region having an amino acid sequence therein of E-P-K-S-S-D-K-T-H-T-S-P-P-S-P (SEQ ID NO:4); and   (2) the carboxy-terminal portion of a human constant region with an additional portion of the constant region, and including a linker between light and heavy chain sequences, the hinge region having the amino acid sequence therein of SEQ ID NO:4 being located between the variable region sequences and the constant region sequences, the single-chain recombinant antibody having complementarity-determining regions identical with those of the single-chain recombinant antibody derived from monoclonal antibody 35.1 and produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277.   
     
     
       11. A single-chain recombinant antibody having specific binding affinity for CD2 a antigen, including therein a human constant region sequence including: (1)a hinge region, the hinge region having an amino acid sequence therein of E-P-K-S-S-D-K-T-H-T-S-P-P-S-P(SEQ ID NO:4); and   (2)the carboxy-terminal portion of a human constant region with an additional portion of the constant region, and including a linker between light and heavy chain sequences, the hinge region having the amino acid sequence therein of SEQ ID NO:4 being located between the variable region sequences and the constant region sequences, the single-chain recombinant antibody being a single-chain recombinant antibody competing with the single-chain recombinant antibody derived from monoclonal antibody 35.1 and produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277 at least about 80% as effectively on a molar basis as the single-chain recombinant antibody derived from monoclonal antibody 35.1 and produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277.   
     
     
       12. The single-chain recombinant antibody of claim 11 wherein the carboxy terminal portion of the human constant region is a sequence from IgG1. 
     
     
       13. The single-chain recombinant antibody of claim 10 wherein the additional portion of the human constant sequence includes the C H  2 and C H  3 regions. 
     
     
       14. The single-chain recombinant antibody of claim 11 wherein the additional portion of the human constant sequence includes the C H  2 and C H  3 regions. 
     
     
       15. The single-chain recombinant antibody of claim 9 labeled with a detectable marker. 
     
     
       16. The single-chain recombinant antibody of claim 15 wherein the detectable marker is selected from the group consisting of enzymes, paramagnetic materials, members of the avidin-biotin specific binding pair, fluorophores, chromophores, chemiluminophores, heavy metals, and radioisotopes. 
     
     
       17. The single-chain recombinant antibody of claim 10 labeled with a detectable marker. 
     
     
       18. The single-chain recombinant antibody of claim 17 wherein the detectable marker is selected from the group consisting of enzymes, paramagnetic materials, members of the avidin-biotin specific binding pair, fluorophores, chromophores, chemiluminophores, heavy metals, and radioisotopes. 
     
     
       19. The single-chain recombinant antibody of claim 11 labeled with a detectable marker. 
     
     
       20. The single-chain recombinant antibody of claim 19 wherein the detectable marker is selected from the group consisting of enzymes, paramagnetic materials, members of the avidin-biotin specific binding pair, fluorophores, chromophores, chemiluminophores, heavy metals, and radioisotopes. 
     
     
       21. A composition comprising: (a) the single-chain recombinant antibody of claim 10 in a quantity sufficient to inhibit production of HIV-1 in infected T cells; and   (b) a carrier.   
     
     
       22. A composition comprising: (a) the single-chain recombinant antibody of claim 11 in a quantity sufficient to inhibit production of HIV-1 in infectcd T cells; and   (b) a carrier.   
     
     
       23. A single-chain recombinant antibody having specific binding affinity for CD2 antigen having complementarity-determining regions identical with those of CD2 SFv-Ig produced by expression of a construct cloned in recombinant Escherichia coli culture ATCC No. 69277, the recombinant antibody further including a hinge region having an amino acid sequence therein of E-P-K-S-S-D-K-T-H-T-S-P-P-S-P (SEQ ID NO:4) and a sufficient portion of the immunoglobulin derived constant region to allow recognition of the Ig-derived amino acid sequence, the hinge region having the amino acid sequence therein of SEO ID NO:4 being located between the variable region sequences and the constant region sequences. 
     
     
       24. A single-chain recombinant antibody having specific binding affinity for CD2 antigen and competing with the single-chain recombinant antibody derived from monoclonal antibody 35.1 and produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277 at least 80% as effectively on a molar basis as the single-chain recombinant antibody derived from monoclonal antibody 35.1 and produced by expression of the construct cloned in recombinant Escherichia coli culture ATCC No. 69277, the recombinant antibody further including a hinge region having an amino acid sequence therein of E-P-K-S-S-D-K-T-H-T-S-P-P-S-P (SEQ ID NO:4) and a sufficient portion of the immunoglobulin derived constant region to allow recognition of the Ig-derived amino acid sequence, the hinge region having the amino acid sequence therein of SEQ ID NO:4 being located between the variable region sequences and the constant region sequences.